JP2021500912A5

JP2021500912A5 -

Info

Publication number: JP2021500912A5
Application number: JP2020524308A
Authority: JP
Filing date: 2018-10-30
Publication date: 2021-12-02
Anticipated expiration: 2038-10-30

Claims

A method for determining the selectivity of a test compound, the following steps:
(A) To provide a sample containing at least two distinguishable subpopulations of cells in a total population of cells;
(B) Divide the sample into at least two parts;
(C) Incubate at least one portion obtained in step (b) in the absence of the test compound and at least one portion obtained in step (b) in the presence of the test compound;
(D)
A total population of cells exhibiting the same phenotype in (i) the at least one portion incubated in the presence of the test compound and (ii) the at least one portion incubated in the absence of the test compound. Determining the number of cells in one of the at least two subpopulations exhibiting a distinguishable phenotype with respect to the number of cells; and (e) determining the selectivity of the test compound (i). ) Is divided by (ii) to derive the phenotype referenced in (d) of said one subgroup referred to in step (d) in all other subpopulations, where (i). is larger than a value obtained by dividing 1 by the (ii), before Symbol test compound is divided by selectively inducing the phenotype referenced (d), the and (i) (ii) If it is less than 1, selectively inhibiting or reducing the phenotype referred to in (d),
Including, how.

In the step (e), when the value obtained by dividing (i) by (ii) is larger than 1.05, 1.1, 1.5, 2, 3 or 5, the test compound is referred to in (d). The method of claim 1, wherein the referenced phenotype is selectively derived.

In the step (e), when the value obtained by dividing (i) by (ii) is less than 0.95, 0.9, 0.7, 0.5, 0.3 or 0.2, the test is performed. The method of claim 1, wherein the compound selectively inhibits or reduces the phenotype referred to in (d).

If the distinguishable phenotype of step (d) is survival, where (i) the selectivity determined in step (e) is less than 1, the test compound is said to be step (d). ) Is determined to selectively reduce the number of viable cells in the one subpopulation, and (ii) if the selectivity determined in step (e) is greater than 1, the test compound is said to be said. Selectively improve the survival rate of one subpopulation and / or selectively reduce the survival rate of one or more of the subpopulations (s) other than the one subpopulation of step (d). The method of claim 1, wherein it is determined.

A method of determining whether a subject suffering from cancer responds to or is responsive to treatment with a test compound, the following steps:
(A) To provide a sample obtained from a subject comprising at least two subpopulations of cells in a total population cell, wherein at least one subpopulation corresponds to a cancerous cell and at least one. Subpopulations correspond to non-cancerous cells;
(B) Divide the sample into at least two parts;
(C) Incubating at least one portion obtained in step (b) in the absence of the test compound and at least one portion in the presence of the test compound;
(D)
To the number of viable cells in the total population of (i) the at least one portion incubated in the presence of the test compound and (ii) the at least one portion incubated in the absence of the test compound. In contrast, the subject is treated with the test compound by determining the number of living cells in at least one of the subpopulations corresponding to the cancer cells; and by dividing (e) (i) by (ii). to is to determine whether there is or responsive reply, wherein said subject, if said obtained value is less than 1, there is or responsive to respond to treatment,
Including, how.

In step (e), the subject responds to or responds to treatment if the resulting value is less than 0.95, 0.9, 0.8, 0.6, 0.4 or 0.2. The method of claim 5, which has the property.

The method is repeated for at least two test compounds, and whether the subject is responsive or responsive to treatment with the combination of the at least two test compounds is from 1.0 to at least each of the two test compounds. The value obtained in (e) is subtracted to obtain the sum of the obtained values for the at least two test compounds, and when the obtained sum is greater than -1 , the subject is a combination of the at least two test compounds. 5. The method of claim 5, wherein the method is determined to be responsive or responsive to treatment with.

The method is repeated for at least two test compounds, and whether the subject is responsive or responsive to treatment with the combination of the at least two test compounds is from 1.0 to at least each of the two test compounds. Subtract the value obtained in (e) to give the sum of the values obtained for the at least two test compounds, and if the sum obtained is greater than -0.5, 0, 0.5 or 1, the subject. 5. The method of claim 5, wherein is determined to be responsive or responsive to treatment with a combination of at least two test compounds.

The method according to any one of claims 1 to 8 , wherein the test compound (s) comprises one or more chemical substances.

At least one moiety obtained in step (b) is further divided into at least two moieties, where each of the at least two moieties is incubated with the test compound at a different concentration in step (c) and step (d). And (e) are independently repeated at each concentration of the test compound to determine the selectivity / value of each concentration of the test compound, whereby the average selectivity / value of all concentrations is determined in the step (e). The method according to any one of claims 1 to 9 , which is calculated after) and used to determine the final selectivity / value.

In step (b), the sample is divided into at least three parts, and in step (c), at least two parts are incubated in the absence of the test compound and / or at least two parts are in the presence of the test compound. Each portion incubated under, thereby incubating in the presence of said test compound, is incubated in the presence of said test compound at the same concentration to give the same distinguishable phenotype in step (d). For the total number of populations of the cells shown, the number of cells in one of the at least two subpopulations showing the distinguishable phenotype is:
(I) Independently determined for each portion incubated in the presence of the test compound and / or (ii) independently incubated in the absence of the test compound, and (i). The method according to any one of claims 1 to 9 , wherein the average of the relative numbers obtained and / or the average of the relative numbers obtained in (ii) is used.

In step (b), the sample is divided into at least three parts, in step (c), at least one part is incubated in the absence of the test compound and / or at least two parts are of the test compound. Incubated in the presence of at least two different concentrations and in step (d) show the distinguishable phenotype with respect to the total population of cells showing the same distinguishable phenotype. The number of cells in one of the at least two subpopulations
(I) Independently determined for each moiety incubated in the presence of the test compound and / or (ii) Independently determined for each moiety incubated in the absence of the test compound, where (ii) The average of i) is determined independently for each concentration and / or the average of said (ii) is determined and used for further steps, wherein in step (e) the selectivity / value is determined. The final selectivity / value is determined by dividing the average of (i) by the average of (ii) for each concentration, and the final selectivity / value is the average of the selectivity / value for each concentration. The method according to any one of claims 1 to 9 , which is obtained by the above-mentioned method.

The method is repeated for at least two test compounds, the test compound having the lowest value obtained in step (e) is selected to treat the subject suffering from cancer, according to claim 5 or 9-12 The method described in any one of the above.

The method is repeated for at least three test compounds, and for each of the at least two test compounds in the combination, the value obtained in (e) is subtracted from 1.0 to obtain the at least two test compounds in the combination. is a combination of at least two of the at least three test compounds with the highest value obtained by adding the value of the result is selected for treatment of said subject suffering from cancer, claims 7 to 12 The method according to any one of the above.

The method according to any one of claims 5 to 14 , wherein the cancer is a cancer associated with PBMC or bone marrow cells or cells derived from PBMC or bone marrow cells.

The method according to any one of claims 1 to 15 , wherein the sample is a tissue sample containing at least 1% cancerous cells and / or at least 1% non-cancerous cells.

The method according to any one of claims 1 to 16 , wherein the tissue sample is cultured as a non-adhesive cell monolayer.

17. The method of claim 17, wherein the viable and non-cancerous cells are determined using an automated microscope.

18. The method of claim 18 , wherein the number of viable cells is determined as the number of unfragmented nuclei.